Spectrophotometric and colorimetric determination of protein concentration.
Spectrophotometric and colorimetric determination of protein concentration.
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DOI：
10.1002/0471142727.mb1001as76
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发表时间：
2006-11-01
影响因子：
--
通讯作者：
Smith, John A
中科院分区：
文献类型：
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作者：
Simonian, Michael H;Smith, John A
本单元描述了用于测量溶液中样品蛋白质浓度的分光光度法和比色法。280 nm处的吸光度测量用于通过与标准曲线或该蛋白质的公布吸光度值进行比较来计算蛋白质浓度。替代方案使用205 nm处的吸光度计算蛋白质浓度。这两种方法都可用于定量粗裂解物和纯化或部分纯化的蛋白质中的总蛋白质。还描述了使用分光荧光计或滤光荧光计来测量样品溶液的固有荧光发射。将测量结果与标准溶液的排放量进行比较，以确定纯化蛋白质的浓度。布拉德福德比色法，基于染料考马斯亮蓝结合到一个未知的蛋白质，还介绍了，因为是劳里方法，它测量酪氨酰残基在一个未知的比色反应。
This unit describes spectrophotometric and colorimetric methods for measuring the concentration of a sample protein in solution. Absorbance measurement at 280 nm is used to calculate protein concentration by comparison with a standard curve or published absorptivity values for that protein. An alternate protocol uses absorbance at 205 nm to calculate the protein concentration. Both methods can be used to quantitate total protein in crude lysates and purified or partially purified protein. Use of a spectrofluorometer or a filter fluorometer to measure the intrinsic fluorescence emission of a sample solution is also described. The measurement is compared with the emissions from standard solutions to determine the concentration of purified protein. The Bradford colorimetric method, based upon binding of the dye Coomassie brilliant blue to an unknown protein, is also presented, as is the Lowry method, which measures colorimetric reaction of tyrosyl residues in an unknown.